ABSTRACT: A congenic mouse line was constructed by introgressing a C3H chromosome 9 region harboring Ath29 into the C57BL/6 apoE-deficient background. RNA was extracted from aorta using a QIAGEN kit . Total RNA was pooled in an equal amount from 3 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in the aorta of Ath29 congenic mice and C57BL/6 apoE-deficient mouse strains fed a chow or western diet. 4 groups of mice were studied: C57BL/6 apoE-/- control mice and Ath29 congenic mice fed a chow or Western diet. After fed a Western diet for 2 weeks, aorta was harvested and total RNA prepared.
Project description:Inbred mouse strains C57BL/6J (B6) and C3H/HeJ (C3H) exhibit a marked difference in their susceptibility to atherosclerosis and the arterial wall has proven to be a source of the difference in atherosclerosis susceptibility. Genome-wide gene expression analysis was conducted in aortic walls of the two strains. Total RNA was extracted from aortas of 6-week-old female B6 and C3H apoE-deficient (apoE-/-) mice fed a chow or Western diet. 1514 genes in chow fed mice and 590 genes in Western fed mice were found to be differentially expressed between the two strains. RNA was extracted from aorta using a Trizol protocol. Total RNA was pooled in an equal amount from 4 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in aortic walls of two apoE-deficient mouse strains when fed a chow or western diet. Experiment Overall Design: 4 groups of mice were studied: C57BL/6 apoE-/- mice on chow diet (03-62_BC), C57BL/6 apoE-/- mice on Western diet (03-62_BW), C3H/HeJ apoE-/- mice on chow diet (03-62_CC), and C3H/HeJ apoE-/- mice on Western diet (03-62_CW). Mice were weaned at 3 weeks of age onto a chow diet. At 4 weeks of age, mice were switched onto a western diet or continued the chow diet for 2 additional weeks.
Project description:A congenic mouse line was constructed by introgressing a C3H chromosome 9 region harboring Ath29 into the C57BL/6 apoE-deficient background. RNA was extracted from aorta using a QIAGEN kit . Total RNA was pooled in an equal amount from 3 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in the aorta of Ath29 congenic mice and C57BL/6 apoE-deficient mouse strains fed a chow or western diet.
Project description:A congenic line was constructed by introgressing a C3H chromosome 1 region harboring Bglu3 into C57BL/6 apoE-/- background. RNA was extracted from liver using a QIAGEN kit . Total RNA was pooled in an equal amount from 3 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in the liver of Bglu3 congenic mice and C57BL/6 apoE-deficient mouse strains fed a western diet. 2 groups of mice were studied: C57BL/6 apoE-/- control mice and Bglu3 congenic mice. After fed a Western diet for 12 weeks, liver was harvested and total RNA prepared.
Project description:Inbred mouse strains C57BL/6J (B6) and C3H/HeJ (C3H) exhibit a marked difference in their susceptibility to atherosclerosis and the arterial wall has proven to be a source of the difference in atherosclerosis susceptibility. Genome-wide gene expression analysis was conducted in aortic walls of the two strains. Total RNA was extracted from aortas of 6-week-old female B6 and C3H apoE-deficient (apoE-/-) mice fed a chow or Western diet. 1514 genes in chow fed mice and 590 genes in Western fed mice were found to be differentially expressed between the two strains. RNA was extracted from aorta using a Trizol protocol. Total RNA was pooled in an equal amount from 4 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in aortic walls of two apoE-deficient mouse strains when fed a chow or western diet. Keywords: atherosclerosis, arterial walls, C57BL/6, C3H/HeJ, Inbred strains, Hyperlipidemia, and Western diet
Project description:Identification of novel pathways in the development of atherosclerosis. Here, we are looking at changes in gene expression that occur in the aorta with the development of atherosclerosis Analysis used RNA from thoracic aortas from chow fed ApoE knockout mice as control samples for comparison to the experimental samples from 8 week and 16 week ApoE knockout mice fed a western-type diet
Project description:SHP (small heterodimer partner; NR0B2) belongs to the nuclear hormone receptor superfamily, which regulates numerous developmental and metabolic cellular functions. To study physiological function of SHP, we generated congenic SHP-/- mice on C57Bl/6 background. When the congenic SHP-/- mice were challenged with a western diet (harlan, TD.88137) for 22 weeks, they were resistant to diet induced obesity and hepatic steatosis compared to WT controls. However, their hepatic insulin sensitivity was compromised when assessed with phospho-Akt levels after insulin injection. Therefore, we investigated hepatic gene expression using illumina beadchip array to explore mechanisms underneath the unique liver physiology in SHP-/- mice. Livers were collected from C57Bl/6 wild type and C57Bl/6 SHP-/- mice fed chow or western diet. The 1 microgram of total RNA obtained from individual mouse (n=4 per group) and subjected to illumina beadchip gene expression profiling.
Project description:A congenic line was constructed by introgressing a C3H chromosome 1 region harboring Bglu3 into C57BL/6 apoE-/- background. RNA was extracted from liver using a QIAGEN kit . Total RNA was pooled in an equal amount from 3 mice for each group. Standard Affymetrix procedures were performed using 8ug of total RNA. Microarrays were used to detect gene expression in the liver of Bglu3 congenic mice and C57BL/6 apoE-deficient mouse strains fed a western diet.
Project description:ApoE-/-mice were fed chow or Western diet for 12 weeks and NPRC expression was significantly increased in the aortic tissues of Western diet-fed mice. Systemic NPRC knockout mice were crossed with ApoE-/- mice to generate ApoE-/-NPRC-/- mice, and NPRC deletion resulted in a significant decrease in the size and instability of aortic atherosclerotic lesions in ApoE-/-NPRC-/- versus ApoE-/- mice.
Project description:A 6-month in vivo study will be carried out at Case Western Reserve University (CWRU) using an ApoE-/- mouse model of atherosclerosis (obtained from The Jackson Laboratory), which will be treated as follows: Control: Mice fed a chow diet (control) (n=12) Progression: Mice fed a high-fat diet with cholesterol (n=12) Regression: Mice fed a high-fat diet with cholesterol followed by a switch to a chow diet (n=12) At the end of the experiment, mice will be euthanized and aorta excised from each animal. They will be immediately incubated in dialyzed serum-containing medium with uniformly C13 labelled glucose for 24 h and then extracted for Stable Isotope Resolved Metabolomics (SIRM) experiments. The incubation media and tissue extracts will be sent to University of Kentucky’s RC-SIRM: Resource Center for Stable Isotope-Resolved Metabolomics for analysis. The samples will undergo the following modes of analysis: nuclear magnetic resonance (NMR), Ion Chromatography Coupled with Mass Spectrometry (ICMS)
Project description:Inflammatory monocyte (MC) subset differentiation is a major feature in tissue inflammatory and atherosclerosis. The underlying molecular mechanism remains unclear. This study aims to explore molecule targets and signaling which determinate immunological features in MC subsets. ApoE-/- mice were fed a normal chow diet and switched to a high-fat (HF) diet at age 8 weeks and maintained on a HF diet for 12-14 weeks. The control C57BL/6 mice were fed a normal chow diet throughout. Blood Ly6Chigh and Ly6Clow MC subsets from control and ApoE-/- mice were isolated by flow cytometry sorting and subjected for bulk high-throughput RNA-sequencing.